State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China.
Department of Anesthesiology, Sir Run Run Hospital Affiliated to Nanjing Medical University, Nanjing 211166, China.
J Nanosci Nanotechnol. 2020 Mar 1;20(3):1332-1340. doi: 10.1166/jnn.2020.17356.
Nucleic acid aptamers are a kind of one-dimensional biological nanomaterials and have found many applications. This paper designed and implemented a polymerase chain reaction (PCR) amplification device with a reaction volume of 500 L, which can be used for the amplification of nucleic acid aptamers of tumor cells in the aptamer selection. This device mainly includes a control module, a temperature measuring module, a PCR amplification tube, a metal tank module, a liquid crystal display (LCD) and operation module and a cooling module. The new PCR amplification chamber is matched with the designed metal tank to ensure the temperature uniformity of the PCR amplification solution. The control module based on the STM32F103RCT6 manages the workflow of the entire device. The PCR amplification chamber and PT100 sensors on the metal tank formed a closed-loop feedback system, and the incremental proportional-integral-derivative (PID) algorithm was used to achieve the precise temperature control. In addition, we introduced the Smith predictive compensation algorithm to solve the temperature hysteresis problem of the PCR amplification chamber. The experimental results showed that the PCR device can meet the requirements for the nucleic acid aptamer selection of tumor cells. The device can also be used in other experiments with large-volume PCR amplification.
核酸适体是一种一维生物纳米材料，具有广泛的应用。本文设计并实现了一种 500 μL 反应体积的聚合酶链反应（PCR）扩增装置，可用于适体筛选过程中肿瘤细胞核酸适体的扩增。该装置主要包括控制模块、测温模块、PCR 扩增管、金属槽模块、液晶显示屏（LCD）和操作模块、以及冷却模块。新型 PCR 扩增室与设计的金属槽相匹配，以确保 PCR 扩增溶液的温度均匀性。基于 STM32F103RCT6 的控制模块管理整个装置的工作流程。金属槽上的 PCR 扩增室和 PT100 传感器形成闭环反馈系统，采用增量式比例-积分-微分（PID）算法实现精确的温度控制。此外，我们引入了史密斯预估补偿算法来解决 PCR 扩增室的温度滞后问题。实验结果表明，该 PCR 装置能够满足肿瘤细胞核酸适体筛选的要求。该装置也可用于其他需要大体积 PCR 扩增的实验。
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